comparative proteomics Essay

This technique separates Rubisco samples based on their size. The electrophoresis has a positive and a negative end. Positive charge proteins are loaded from the positive end and migrate towards the negative end. Negative charge proteins are loaded from the negative end and migrate towards the positive end (Sakthivel & Palani, 2016). The sample that contained the highest molecular weight of Rubisco will travel the shortest distance on the gel while the protein with the smallest molecular weight will travel the longest distance (Sakthivel & Palani, 2016). The size proportion of each Rubisco molecule correlates with the distance traveled. Rubisco will be in its purest form after running through SDS-page since each technique will increase the purity of the protein. If the salting out, the ion exchange and the SDS-page protein isolation techniques are performed on protein Rubisco, then it is purified and separated by solubility, charge, and size. The rationale of this experiment is to isolate the purest form of Rubisco so that it can perform carbon fixation at an optimal

Figure 1 contains gel electrophoresis for protein samples. The lanes were labeled from 1 to 10 from the right to the left. Lane 1 contained the ladder fragment. Lane 2 contained the filtrate. Lane 3 contained the S1 sample. Lane 4 contained the P1 sample. Lane 5 contained the P1 medium salt sample. Lane 6 contained the P1 high salt sample. Lane 7 contained the S2 sample. Lane 8 contained the P2 sample. Lane 9 contained the P2 medium salt sample. Lane 10 contained the P2 high salt sample.

c. Describe the information you can deduce by comparing and contrasting the positive control lane and lane 2 protein bands. By comparing lane 3 the positive control to lane 2 the purified protein X it is noticeable that the first band is the unknown protein X due to the fact that the positive control only contains purified protein X and only forms one band which matches up with the first band in lane 2.

Gel electrophoresis method. Qualitative analysis shows protein concentrations in kidney, heart, and liver. 1-6 are kidney tissue. 7-14 are liver and 15-19 are the heart tissue.

Another machine that is used is the Molecular Dynamics Fluorimager FSI. This device is a quantitative gel and blot system in order for scientists to read. This labels the DNA and proteins on micro plates. A laser scans the DNA a then provides an image for studying the DNA (Harkey, 2002).

ELISA works on the principle of an antigen binding to specific antibody (lock and key), which can be used as a way to identify quantities of proteins in a small sample of fluid. The specific proteins used in an ELISA are estimated quantitatively. The ELISA test is carried out by incubating the serum that contains the antigen of interest with antibody’s within a well, in order for the antibody’s to bind with the specific antigens. The plate is then washed with a mild detergent in order to remove any proteins that have not been bound. The washing of the plates is carried out between every step in order

These steps will repeat for every enzyme concentration’s control, substrate and enzyme test tubes. The total amount of data points should be 6, 2 data points per concentration. After part one, clean the test tubes by correctly disposing of the substances in the waste bin under the fume hood and then proceed to clean with soap so that the test tubes can be used for part

The purpose of this experiment is to perform a procedure known as SDS-PAGE (sodium dodecyl sulfate - polyacrylamide gel electrophoresis) in order to determine protein profiles for seven fish species: Yellow Tuna (Thunnus albacares), Thresher Shark (Alopias vulpinus), Paiche (Arapaima gigas), Swordfish (Xiphias gladius), Catfish (Ameiurus catus), Coho Salmon (Oncorhynchus kisutch), and Rainbow Trout (Oncorhynchus mykiss). The gel electrophoresis separates the fish muscle proteins according to their molecular weights, which allows one to determine a profile for each specie.

We turned the Spectrophotometer on and set the value for absorbance to 475nm. We allowed it to properly warm up for at least 15 minutes before we started using it. We prepared 8 clean test tubes to make reaction assay tubes with 4mL of buffer and .5ml of the enzyme in each tube. They were labeled 1 to 8 starting from the lowest concentration to the highest. Tubes 1,3,5 & 7 had .5 ml of buffer added instead to act as the control group.

Once the protein goes through the pores of the gel based on negative charge, if the charge is high, then the faster it moves in an electric field. The shape is important to know based on shape how the protein would travels through it, for example a round shape will make the protein move faster than rod shape due to the rod will bend which this will cause the protein to stop. The size of the is important to determine whether its big or small if its big then the protein are able to have difficulties to get through the pores and move slowly. The protein travel differently than other natural protein in the gel. The mutation change impacts the negatively charge amino acid with hydrophobic amino acid, which cause the protein to change the charge and shape. This process is to either introduce or eliminate a restriction enzyme, which allows it to eliminate the pieces of DNA whether they have it or not. Each of the restriction enzymes is able to identify and eliminate it, which cause the DNA not a normal DNA because of the size of fragments that can be apart from using gel electrophoresis. This causes the DNA fingerprint to change by different pattern on the

Traditional reagent strip testing for protein uses the principle of the protein error of indicators to produce a visible colorimetric reaction. The protein (primarily albumin) accepts hydrogen ions from the indicator causing change in color. (Susan King Strasinger, 2008)

Question 3: Tuna and Cod will be the most related animals. They both swim and migrate in large schools and can be found in the Pacific Ocean. They both have multiple dorsal fins and can be found in a deeper level of the ocean. This indicates that their protein profiles could be similar.

Agarose gel electrophoresis is a method of gel electrophoresis used in biochemistry, molecular biology, genetics, and clinical chemistry to separate a mixed population of DNA or proteins in a matrix ofagarose. The proteins may be separated by charge and/or size

The last part of this procedure, antibodies will be used to detect one specific protein from others on the membrane. Incubate the membrane with 10 ml of primary antibody for 10-20 minutes and then place on a rocking platform. Rinse the membrane quickly after with wash buffer on a rocking platform. After three minutes is up, discard the wash and incubate the membrane with 10 ml of secondary antibody for 5-15 minutes on the platform. Rinse the membrane again shortly after and wash the membrane for three minutes. Next, discard the wash and add 10 mL of HRP color detection reagent. Incubation will occur after this for 10-30 minutes. Once it is done, rinse the membrane twice with distilled water and blot dry.

For protein portrayal mass spectrometry (MS) uses mass investigation. Although DNA microarray innovation is taking into account an exceptionally delicate and particular hybridization response between nucleic acid sections, basic constraints of biological MS (1) require a few separate ways to deal with protein examination. Usage of the procedures such as test planning, front-end partition, ionization, information procurement, and data analysis that contrasts upon relying the specimen intricacy and examination of the objectives (2).